(a) (b) (c) Figure 1: Bacterial plates after overnight incubation at 37°C. (a) BL21 cells transformed with gbr22 on an Agar Amp plate, (b) BL21 cells with no plasmid DNA on an Agar Amp plate, and (c) the fun plate with bacteria from my shoe with no plasmid DNA on an Agar plate without Amp.
Figure 2: Bacterial culture (BL21+gbr22+LBbroth+amp) culture after 24 hours in the shaking incubator at 37°C and 200-350 rpm.
Figure 3: Cell pellet after bacterial culture from Figure 2 was centrifuged at 4°C and 5,000 rpm for 10 minutes and the liquid was decanted.
Figure 4: Elution 1 (~5mL) and Elution 2 (~5mL) 250 mM Imidazole buffers after being allowed to flow through the column containing the purple gbr22 protein bound to the Ni-NTA.
Figure 5: NanoDrop absorption spectra for Elution 1 at 280 nm (left) and at 574 nm (right).
Figure 6: NanoDrop absorption spectra for Elution 2 at 280 nm (left) and at 574 nm (right).
Figure 7: SDS-PAGE gel after being run for 25 minutes at 200 V in 500 mL 1x TGS buffer and dried at 75 degrees Celsius for 90 minutes. 1=ColorPlus ladder; 2=Cell Lysate (Sample 1); 3=Soluble Faction (Sample 2); 4=Waste Flow Through (Sample 3); 5=Wash with 5mL 20mM Imidazole (Sample 4); 6=Elution 1 Flow through of 5mL 250mM Imidazole (Sample 5); 7=Elution 2 Flow Through of 5mL 250 mM Imidazole (Sample 6); 8=Elution 1 (Jairo); 9=Elution 2 (Jairo).
Figure 8: Molecular Weight Standards for the ColorPlus Ladder
Introduction:
Materials & Methods:
Results:
(a) (b) (c)
Figure 1: Bacterial plates after overnight incubation at 37°C. (a) BL21 cells transformed with gbr22 on an Agar Amp plate, (b) BL21 cells with no plasmid DNA on an Agar Amp plate, and (c) the fun plate with bacteria from my shoe with no plasmid DNA on an Agar plate without Amp.
Figure 2: Bacterial culture (BL21+gbr22+LBbroth+amp) culture after 24 hours in the shaking incubator at 37°C and 200-350 rpm.
Figure 3: Cell pellet after bacterial culture from Figure 2 was centrifuged at 4°C and 5,000 rpm for 10 minutes and the liquid was decanted.
Figure 4: Elution 1 (~5mL) and Elution 2 (~5mL) 250 mM Imidazole buffers after being allowed to flow through the column containing the purple gbr22 protein bound to the Ni-NTA.
Figure 5: NanoDrop absorption spectra for Elution 1 at 280 nm (left) and at 574 nm (right).
Figure 6: NanoDrop absorption spectra for Elution 2 at 280 nm (left) and at 574 nm (right).
Figure 7: SDS-PAGE gel after being run for 25 minutes at 200 V in 500 mL 1x TGS buffer and dried at 75 degrees Celsius for 90 minutes. 1=ColorPlus ladder; 2=Cell Lysate (Sample 1); 3=Soluble Faction (Sample 2); 4=Waste Flow Through (Sample 3); 5=Wash with 5mL 20mM Imidazole (Sample 4); 6=Elution 1 Flow through of 5mL 250mM Imidazole (Sample 5); 7=Elution 2 Flow Through of 5mL 250 mM Imidazole (Sample 6); 8=Elution 1 (Jairo); 9=Elution 2 (Jairo).
Figure 8: Molecular Weight Standards for the ColorPlus Ladder
Discussion:
Conclusions:
References: